The zebra fish has emerged as a powerful model system to study neural development, physiology, and disease. It is ideal for this particular purpose since it is a small and translucent vertebrate with a wide behavioral repertoire. Despite this progress, there are still comparatively few transgenic lines that allow the full exploitation f optogenetic approaches. For example, the field still lacks specific drivers and reporters for most neurotransmitters or peptides, and existing lines that mark specific brain regions or nuclei often lack the necessary specificity. The zebra fish as a model system is thus still lacking the essential tool set that the Drosophila community has built in the last decade. To overcome these limitations, we propose to generate transgenic lines that allow the specific expression of proteins of interest in restricted subpopulations of neurons that are specified by brain region and/or by neurotransmitter subtype. We will use enhancer trapping to generate 240 lines that drive region- and cell- type specific neural expression via a modified Gal4 vector (Aim 1), and we will use genome engineering to generate 12 lines that drive expression in neurons producing the major classes of neurotransmitters (Aim 2). An essential component of the enterprise will be the design and establishment of an online interface that will provide details of each transgenic line. Researchers will be provided with extensive details of each line's characteristics and well-annotated image datasets. The Engert and Schier lab have extensive expertise in anatomical and functional imaging of the zebra fish nervous system as well as the molecular and genetic toolbox that is necessary for the proposed project. The project will be further supported by an advisory committee consisting of leaders in the field of zebra fish neurobiology.